In vivo validation of bioprinted gingival biografts based on mesenchymal cells in hyaluronic acid scaffold

• Grant number: 21/12069-1
• Support Opportunities: Research Grants - Innovative Research in Small Business - PIPE
• Start date: March 01, 2022
• End date: July 31, 2024
• Field of knowledge: Health Sciences - Dentistry - Periodontology
• Agreement: SEBRAE-SP
• Principal Investigator: Juliana Magro Ribeiro
• Grantee: Juliana Magro Ribeiro
• Company: In Situ Terapia Celular Ltda
• CNAE: Pesquisa e desenvolvimento experimental em ciências físicas e naturais
• City: Ribeirão Preto
• Associated researchers: Adriana Oliveira Manfiolli ; Carolina Caliári Oliveira ; Francisco Jean Pierre Romero Febres ; Mario Taba Junior ; Valessa Florindo Carvalho
• Associated research grant: 20/05894-3 - Technical validation of the construction of a gingival bioprinted reconstructor based on mesenchymal cells or extracellular vesicles in hyaluronic acid scaffold, AP.PIPE
• Associated research grant(s): 21/14943-0 - In vivo validation of bioprinted gingival biografts based on mesenchymal cells in hyaluronic acid scaffold, AP.PIPE
• Associated scholarship(s): 23/02445-1 - In vivo validation of bioprinted gingival biograft based on mesenchymal cells on hyaluronic acid scaffold., BP.TT; 22/07988-0 - In vivo validation of bioprinted gingival biograft based on mesenchymal cells on hyaluronic acid scaffold, BP.TT; 22/06240-2 - In vivo validation of bioprinted gingival biograft based on mesenchymal cells on hyaluronic acid scaffold, BP.TT; 22/03560-6 - In vivo validation of bioprinted gingival biograft based on mesenchymal cells on hyaluronic acid scaffold, BP.PIPE

Abstract

Gingival recession is a condition characterized by oral exposure of the root surface due to a displacement of the apical gingival margin to the cementoenamel junction. The most common consequences of gingival recession are deterioration of dental esthetics, as well as dentin hypersensitivity and even tooth loss. Currently, one of the most used procedures for root coverage is the one using connective tissue graft plus coronally positioned flap (CTG + CAF). However, surgeries that involve graft harvesting are time-consuming and may have high morbidity, due to the need for a second surgical site, in addition to increasing patient discomfort, post-surgical bleeding, and limited supply of donor tissue. For these reasons, alternative surgical techniques to CTG + CAF are proposed, as well as the use of some products to reduce gingival recessions. Unlike all products found on the market, we developed a biograft based on cell therapy that is composed of a scaffold of hyaluronic acid (HA) and that contains mesenchymal cells (CM) incorporated. This biotint is bioprinted on a 3D bioprinter which gives scalability to production that is totally based on the GMP (Good Manufacturing Practice) concept. During the first phase of biograft development, the protocol for obtaining the HA-based biomaterial was established, which presented texture and bioprinting and handling conditions. Once the conditions of the biomaterial were determined, it was demonstrated that this scaffold and the bioprinting process do not affect the viability of the contained CMs. In addition, it was verified in vitro that the biograft is not cytotoxic to cells belonging to the oral mucosa such as gingival keratinocytes and that it promotes the proliferation of human gingival fibroblasts (FGH) in a dose-dependent manner, therefore, the biograft with 1.5x105 CMs which have the greatest capacity to stimulate the multiplication of FGHs. Added to the proliferative effect of the biograft, we verified in vitro that this product is also capable of promoting greater FGH migration. Once the amount of CMs to obtain the biological results was determined and that favorable biological properties were demonstrated during the proof of concept, further studies are needed to verify the therapeutic effect of biografts in the treatment of gingival recession models in vivo , focusing the analyzes on the parameters required by ANVISA for the release of tests in humans in the future. In addition, also aiming at the entry of this product on the market, it is necessary to establish the proper conditions for storage and transport of biografts so that they are configured as viable products. (AU)