Assessment of fresh and preserved amniotic membrane for guided bone regeneration in mice

Abstract

Thanks to its biological properties, the human amniotic membrane (HAM) can be used as a barrier membrane for guided bone regeneration (GBR). However, no study has assessed the influence of the preservation method of HAM for this application. This study aimed to establish the most suitable preservation method of HAM for GBR. Fresh (F), cryopreserved (C) lyophilized (L), and decellularized and lyophilized (DL) HAM were compared. The impact of preservation methods on collagen and glycosaminoglycans (GAG) content was evaluated using Masson's trichrome and alcian blue staining. Their suture retention strengths were assessed. In vitro, the osteogenic potential of human bone marrow mesenchymal stromal cells (hBMSCs) cultured on the four HAMs was evaluated using alkaline phosphatase staining and alizarin red quantification assay. In vivo, the effectiveness of fresh and preserved HAMs for GBR was assessed in a mice diaphyseal bone defect after 1 week or 1 month healing. Micro‐CT and histomorphometric analysis were performed. The major structural components of HAM (collagen and GAG) were preserved whatever the preservation method used. The tearing strength of DL‐HAM was significantly higher. In vitro, hBMSCs seeded on DL‐HAM displayed a stronger ALP staining, and alizarin red staining quantification was significantly higher at Day 14. In vivo, L‐HAM and DL‐HAM significantly enhanced early bone regeneration. One month after the surgery, only DL‐HAM slightly promoted bone regeneration. Several preserving methods of HAM have been studied for bone regeneration. Here, we have demonstrated that DL‐HAM achieved the most promising results for GBR.

Surgical Application of Human Amniotic Membrane and Amnion-Chorion Membrane in the Oral Cavity and Efficacy Evaluation: Corollary With Ophthalmological and Wound Healing Experiences

“…Gurinsky (2009) reported the ability of processed dehydrated amnion to self-adhere, reducing surgical time and eliminating the need for sutures in the management of gingival recession. Originally designed for ophthalmological use, their current indications have also been extended to wound healing, including diabetic foot, venous leg ulcers and lower third nasal reconstruction (Fetterolf and Snyder, 2012;Koob et al, 2016;Laurent I. et al, 2017;Lullove, 2017;Xue et al, 2018) (Fenelon et al, 2019;Fenelon et al, 2020). We reported in in vivo studies that F-hAM and D-hAM were significantly stronger than C-hAM and L-hAM.…”

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

“…Results suggested that the use of cryopreserved AM with the mesenchymal side in contact with the calvarial defect was the best condition to favor bone regeneration in mice (Fénelon et al, 2018b ). The other study compared four commonly used preservation methods of AM (i.e., fresh, cryopreserved, lyophilized or decellularized, and lyophilized) for GBR procedures in mice tibial defect (Fenelon et al, 2020 ). Covering the defect with lyophilized or decellularized and lyophilized AM significantly enhanced early bone formation.…”

“…The wide heterogeneity of animal models (species, localization and size of the defect) increases the difficulty of comparing these conditioning methods. One study compared four commonly used preservation methods of AM (i.e., fresh, cryopreserved, lyophilized, and decellularized-lyophilized AM) using the same animal diaphyseal bone defect model in mice (Fenelon et al, 2020 ). No significant difference was observed between the empty defect and the defect covered either by fresh or cryopreserved AM, whereas lyophilized and decellularized-lyophilized AM significantly enhanced early bone regeneration.…”

Insights on the Human Amniotic Membrane in Clinical Practice with a Focus on the New Applications in Retinal Surgery

“…The hAM controls the scarring formation through the reduction of fibrosis [34]. This effect is achieved by the downregulation of TGF-b and its receptor expression [35]. The hAM has been used in different surgical subspecialties; it can be used as a dressing or as a transplanted graft.…”