Periodontal ligament cellular structures engineered with electrospun poly(DL‐lactide‐co‐glycolide) nanofibrous membrane scaffolds

Abstract

Periodontal tissue engineering is expected to overcome the limitations associated with the existing regenerative techniques for the treatment of periodontal defects involving alveolar bone, cementum, and periodontal ligament. Cell-based tissue engineering approaches involve the utilization of in vitro expanded cells with regenerative capacity and their delivery to the appropriate sites via biomaterial scaffolds. The aim of this study was to establish living periodontal ligament cell-containing structures on electrospun poly(DL-lactic-co-glycolic acid) (PLGA) nanofiber membrane scaffolds, assess their viability and characteristics, and engineer multilayered structures amenable to easy handling. Human periodontal ligament (hPDL) cells were expanded in explant culture and then characterized morphologically and immunohistochemically. PLGA nanofiber membranes were prepared by the electrospinning process; mechanical tensile properties were determined, surface topography, nanofiber size, and porosity status were investigated with SEM. Cells were seeded on the membranes at approximately 50,000 cell/cm(2) and cultured for 21 days either in expansion or in osteogenic induction medium. Cell adhesion and viability were demonstrated using SEM and MTT, respectively, and osteogenic differentiation was determined with IHC and immunohistomorphometric evaluation of osteopontin, osteocalcin, and bone sialoprotein marker expression. At days 3, 6, 9, and 12 additional cell/membrane layers were deposited on the existing ones and multilayered hybrid structures were established. Results indicate the feasibility of periodontal ligament cell-containing tissue-like structures engineering with PDL cells and electrospun nanofiber PLGA scaffolds supporting cell adhesion, viability and osteogenic differentiation properties of cells in hybrid structures amenable to macroscopic handling.

Engineering on the Straight and Narrow: The Mechanics of Nanofibrous Assemblies for Fiber-Reinforced Tissue Regeneration

“…124 Others have investigated multicomponent scaffolds, where a gelatinous core is established (by injection) in the mid-substance of a nanofibrous sheet for intervertebral disc applications. 125 Still others have begun to investigate how layers of nanofibers can be seeded with cells, and then stacked to achieve anatomic forms such as periodontal ligament 126 as well as more complicated structures recreating the anatomic form of the IVD and the knee meniscus 127,128 (Fig. 15).…”

Advanced drug delivery approaches against periodontitis

“…At 3, 6, 9, and 12 days, it was observed that multilayered hybrid arrangements had developed by deposition of supplementary membrane layers on the existing ones. Thus cell adhesion, viability and osteogenic differentiation was obtained indicating the feasibility of use in periodontal tissue regeneration (Inanç et al, 2009). Kearns and coworkers coated smooth glass and rough titanium substrates with polytetrafluoroethylene (PTFE) nanofibers, cultured human gingival fibroblasts on the substrates and tested the orientation and collagen deposition.…”

Regeneração periodontal em cães

“…Entretanto, essa técnica caminha agora para outro estágio, no qual as membranas serão multifuncionais e não apenas com a função de estabilizar o defeito (INANÇ et al, 2009;PARK et al, 2009).…”

“…Essas novas membranas estão sendo chamadas multifuncionais, porque sua estrutura física e constituição química são desenvolvidas para apresentar Ciência Rural, v.41, n.12, dez, 2011. alguma das funções: oclusão da borda superior do defeito para impedir a migração do epitélio juncional na direção apical; maior adesão da gengiva à sua superfície externa, diminuindo assim a possibilidade de ocorrer recessão gengival; propriedades mecânicas adequadas à manutenção do espaço e estabilização da matriz provisória; favorecimento da osteocondução; além de serem reabsorvíveis (CHO et al, 2009;INANÇ et al, 2009;PARK et al, 2009).…”

“…Uma opção é a engenharia de tecidos com o cultivo de células diretamente sobre a membrana a ser aplicada sobre o defeito periodontal, como em estudo que analisou a obtenção de células do ligamento periodontal e sua cultura em membranas de nanofibras de PGLA (INANÇ et al, 2009). Outra abordagem é o preenchimento do defeito periodontal pela aplicação direta de células-tronco mesenquimais, como nos recentes estudos de WEI et al (2010) e LIU et al (2010.…”