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Item Analysis of Movements in Spinal Cord Hemisection Treatment with Amniotic Membrane – Preclinical Study(Scientific Research Publishing Inc.) Neves, Marcele Florêncio das; Fonseca, Joyce Lilian Reis da; Carvalho, Paula Cristina Simões de; Pereira, Aline Priscila Campos; Polisel, Evandro Henrique Pintor; Proglhof, Stephanie de Medeiros Passos; Sant ́Anna, Luciana Barros; Lima, Mário Oliveira; Arisawa, Emília Ângela Lo SchiavoAims: This study aimed to evaluate the efficacy of Amniotic Membrane application in rats with Spinal Cord Injury induced by transverse hemisection using kinematic analysis and Sciatic Functional Index. Study design: True experimental research design. Place and Duration of Study: Instiuto de Pesquisa e Desenvolvimento (IP&D) of Universidade do Vale do Paraíba (UNIVAP), between September 2016 and December 2017. Methodology: Fifteen adult male rats were used, allocated into three equal groups: Control (the spinal cord injury and Amniotic Membrane application were simulated), Lesion (spinal cord injury not treated), Amniotic membrane (spinal cord injury treated by amniotic membrane). All animals underwent surgical procedures. A transverse hemisection was performed in groups Lesion and Amniotic Membrene. A fragment of the biomaterial was applied in group AM covering the hemisection area. Results: Sciatic Functional Index and motion analysis were performed by comparing images taken at pre- and postoperative time at 7, 14, 21, and 28 days. The kinematic analysis showed a significant difference between groups Control and Lesion at 7 days (p = 0.023) and 14 days (p = 0.015), and between groups Lesion and Amniotic Membrane at 14 days (p = 0.039), comparing the postoperative periods. The Sciatic Functional Index revealed significant differences between Groups Control and Lesion at 7 (p = -0.002), 14 (p = 0.003), and 21 days (p = 0.009), between Groups Control and Amniotic Membrane at 7 (p = 0.014), 14 (p = 0.007), and 28 days (p = 0; 013), and between Groups Lesion and Amniotic Membrane only at 14 days (p = 0.039). Conclusion: Application of amniotic membrane in spinal cord hemisection in rats induced gait recovery and improvement in SFI compared to the untreated group.Item Antimicrobial Effect of the Amniotic Membrane Isolated and Associated with Photodynamic Therapy(MDPI) Santos, Amanda Cerquearo Rodrigues dos; Teodoro, Guilherme Rodrigues; Ferreira-Strixino, Juliana; Sant’Anna, Luciana BarrosMicrobial control through alternative therapies, such as the amniotic membrane (AM) and antimicrobial photodynamic therapy (aPDT), has been gaining prominence with the advancement of bacterial resistance to conventional treatments. This study aimed to evaluate the antimicrobial effect of AM isolated and associated with aPDT using the PHTALOX® as a photosensitizer (PS) against Staphylococcus aureus and Pseudomonas aeruginosa biofilms. The groups studied were: C+; L; AM; AM+L; AM+PHTX; and AM+aPDT. The irradiation parameters were 660 nm, 50 J.cm−2, and 30 mW.cm−2. Two independent microbiological experiments were carried out in triplicate, and the results were analyzed by CFU/mL counting and a metabolic activity test, both statistically analyzed (p < 0.05). The integrity of the AM was verified after the treatments by a scanning electron microscope (SEM). The groups AM, AM+PHTX, and, mainly, AM+aPDT showed a statistical difference When compared to C+ regarding the decrease in CFU/mL and metabolic activity. SEM analysis showed significant morphological alterations in the AM+PHTX and AM+aPDT groups. The treatments with AM isolated or associated with PHTALOX® were adequate. The association had potentiated the biofilm effect, and the morphological differences presented by AM after treatment did not hinder its antimicrobial effect, encouraging its use in biofilm formation locals.Item Effects of Preservation Methods in the Composition of the Placental and Reflected Regions of the Human Amniotic Membrane(Karger) Moraes, Jéssica Tereza Guedes de Oliveira; Costa, Maíra Maftoum; Alves, Paula Cristina Santos; Sant’Anna, Luciana BarrosThe human amniotic membrane (AM) is emerging as an in- teresting biomaterial for regenerative medicine due to its biological and mechanical proprieties. The beneficial effects of the AM are probably related to its bioactive factors pro- duced by local cells and stored in the stromal matrix. How- ever, the search for a preservation method capable of pre- serving AM properties remains a challenge. The aim of this study was to evaluate important features of 2 anatomical re- gions of the human AM (reflected and placental amnion) af- ter different preservation methods. For this purpose, human placentas were harvested and processed for AM isolation and storage at 2 different conditions: room temperature for 18 h in DMEM (fresh AM) and −80°C in DMEM/glycerol solu- tion for 30 days (cryopreserved AM). After the storage peri- od, the structural integrity of the membrane was assessed by histological and Picrosirius polarization analysis, cellular vi- ability analysis was performed using the MTT assay, and the soluble proteins were quantified with the Qubit Protein Assay Kit. Both preservation protocols reduced the cell viabili- ty, mainly in the placental amnion region of the AM, but pre- served the morphology of epithelial and stromal layers, as well as the organization and distribution of collagen fibers. There was a reduction in soluble proteins only in fresh AM. Importantly, the cryopreserved AM group presented the same concentration as the control group. In conclusion, the cryopreservation using DMEM/glycerol was ideal for pre- serving the structural integrity and soluble protein content, indicating the feasibility of this method in preserving AM for its use in regenerative medicine.