Proliferation and Morphological Assessment of Human Periodontal Ligament Fibroblast towards Bovine Pericardium Membranes: An In Vitro Study

Abstract

Over the past decade regenerative branches of dentistry have taken on more and more importance, resulting in the development of performing scaffold materials. These should induce cell adhesion, support, and guide the tissues’ growth. Among the developed materials, we can include resorbable or non-membranes. The purpose of this study was to investigate the proliferation abilities and the attachment of human periodontal ligament fibroblasts (HPLIFs) over two bovine pericardium membranes with different thicknesses, 0.2 mm and 0.4 mm, respectively. These membranes have been decellularized by the manufacturer, preserving the three-dimensional collagen’s structure. The HPLFs were cultured in standard conditions and exposed to the tested materials. XTT was performed to assess cell proliferation, while light microscopy (LM) and scanning electron microscopy (SEM) observations assessed fibroblast morphology at different times (T1, T2, and T3). Proliferation assays have shown a statistically significant difference in growth at T1 (p < 0.05) in the cells cultured with a thicker membrane compared to the thinner one. LM analysis showed healthy fibroblasts in contact with the membranes, appearing larger and with a polygonal shape. SEM observation demonstrated thickening of the fibroblasts which continued to adhere to the membrane’s surface, with enlarged polygonal shape and developed filipodia and lamellipodia. These results showed a similar cell behavior over the two bovine pericardium membranes, demonstrating a cellular migration along and within the layers of the membrane, binding with membrane fibers by means of filopodial extensions. Knowledge of the effects of the collagen membranes derived from bovine pericardium on cellular behavior will help clinicians choose the type of scaffolds according to the required clinical situation.

Advances Focusing on the Application of Decellularized Extracellular Matrix in Periodontal Regeneration

“…Serena Bianchi et al introduced the decellularized bovine pericardium as a membrane material loading PDLSCs [ 48 ]. The decellularized bovine pericardiums retained the three-dimensional collagen structure for stem cell infiltration.…”

“…hPDL fibroblasts were seeded on the decellularized bovine pericardium and were healthy, large, and polygonal with filopodia and lamellipodia. Other results show that cells migrate along and within the membrane layer, binding to membrane fibers by filamentous extension, showing that decellularized bovine pericardium could serve as a carrier for PDL fibroblasts [ 48 ].…”

“…The fabrication and in vitro analysis of dECM (solid dECM and soluble dECM) of various tissue origins are prerequisites for applications. The dECM can be generated and improved by a variety of methods, and complex periodontal regeneration modules based on the dECM were recently developed [ 20 , 48 , 55 , 87 ].…”

Recent Advances in Functionalized Electrospun Membranes for Periodontal Regeneration

“…They are made of several kinds of biomaterials, a class of materials “designed to take a form that can direct, through interactions with living systems, the course of any therapeutic or diagnostic procedure” [ 63 ]. In this sense, biomaterials can regulate molecular signals and cellular behaviors to achieve tissue regeneration even if they are not loaded with cells or drugs [ 64 ]. They are characterized by unique chemical, mechanical and biological properties; for example, in this case, osteoinductivity and osteoconductivity, which make them suitable and safe to interact with living tissues [ 65 ].…”

Tomographic Evaluation of Alveolar Ridge Preservation Using Bone Substitutes and Collagen Membranes—A Retrospective Pilot Study

“…Grafting materials have specific biological, chemical and mechanical properties, making them suitable for use in humans. The main hallmark of these biomaterials, which are biocompatibility, degradability, adhesion, osteogenic properties, induction properties and low immunogenicity, allow them to be utilised in regenerative processes [ 13 ]. ARP often involves resorbable or non-resorbable barrier membranes which maintain the space for bone infiltration.…”